| Summary: | - 5 - ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Martin Mžik Supervisor: Doc. RNDr. Lenka Skálová PhD. Title of diploma thesis: Determination of estradioldehydrogenases and estronreductates activity Estrogens belong to the most important factors which affect the formation and growth of breast tissue cancer. The most potent estrogen estradiol which is responsible for development of breast cancer is over-synthesized mainly by 17β-hydroxysteroid dehydrogenases (17β-HSDs) in cancer cells. Determination of estrone redustase activity makes possible to get knowledge about enzyme kinetics, which we can use to study influence of potential inhibitors. The aim of diploma thesis was to find acceptable analytical method for determination of estrone reductase and estradiol dehydrogenase activity in subcelluar fraction of cancer cells. Spectrofluorimetric method, based on continuous monitoring of NADPH/NADP+ transformation, as well as chromatographic method (HPLC) based on direct measurement of estrogens by fluorimetric detector have been tested. Experimental data revealed that spectrofluorimetric method is unusable, but liquid chromatography can be successfully used for determination of estrone reductase and estradiol dehydrogenase activity....
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